Simple two-step purification and characterisation of peroxidase from Citrullus colocynthis

Peroxidase is a biotechnologically important enzyme. The purification of peroxidase from the root of Citrullus colocynthis was carried out in a simple two-step process with maximum purity level. The sample was extracted in a high salt buffer, and the enzyme was partially purified with a Q-Sepharose anion exchange column. Final purification was carried out with HighLoad 16/600 Superdex G-75 column. The purified protein was analysed with SDS gel electrophoresis, which suggested a single band of approximately 35 kDa. Further, the enzyme was identified with the help of Mass spectrometric analysis using an ESI-QTOF Mass spectrometer. The study will be helpful for the isolation and its commercial uses in biotechnology.

Automated summary

The purification of peroxidase enzyme from Citrullus colocynthis root was successfully achieved in a two-step process, resulting in maximum purity. The enzyme was extracted in a high salt buffer and partially purified using a Q-Sepharose anion exchange column, followed by final purification with a HighLoad 16/600 Superdex G-75 column. SDS gel electrophoresis confirmed the presence of a single band at approximately 35 kDa, and mass spectrometric analysis using an ESI-QTOF spectrometer further identified the enzyme. This study will contribute to the isolation and commercial applications of this enzyme in biotechnology.